LOCUS       sequence-105628-        2990 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Encodes RVD positions 13-15 in TALEN assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2990)
  AUTHORS   Ma AC, McNulty MS, Tanya PL, Jarryd CM, Martinez-Galvez G, Argue DP,
            Lee HB, Urban MD, Bullard CE, Blackburn PR, Man TK, Clark KJ, Ekker 
            SC
  TITLE     FusX: A rapid one-step TALE assembly system for genome science.
  JOURNAL   Hum Gene Ther. 2016 Jun;27(6):451-63. doi: 10.1089/hum.2015.172
  PUBMED    26854857
REFERENCE   2  (bases 1 to 2990)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1089/hum.2015"; 
            journalName: "Hum Gene Ther"; date: "2016-06"; volume: "27"; issue: 
            "6"; pages: "451-63"
COMMENT     Sequence Label: pFusB3-36
FEATURES             Location/Qualifiers
     source          1..2990
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(3..341)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDSH
                     HLVRGQAMR"
     primer_bind     206..222
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     232..251
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        232..250
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(298..318)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(298..316)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(337..353)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(337..353)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(350..372)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    361..377
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(385..391,392..409,410..415))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    430..451
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             897..1688
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1781..2369
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2270..2289
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2523..2540
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2699..2726)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2818..2904)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2968..2984
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgtctcatgg cctgaccccg gaccaagtgg tggctatcga gaccggcgcc gctacagggc
       61 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      121 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
      181 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac
      241 tcactatagg gcgaattggg taccgggccc cccctcgagg tcctccagct tttgttccct
      301 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
      361 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
      421 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
      481 ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgagacgtct
      541 agaccagcca ggacagaaat gcctcgactt cgctgctacc caaggttgcc gggtgacgca
      601 caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg gttcgtaagc
      661 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
      721 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt
      781 ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat
      841 ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacattatga
      901 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc
      961 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
     1021 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
     1081 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga
     1141 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
     1201 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct
     1261 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
     1321 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
     1381 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
     1441 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
     1501 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
     1561 ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc
     1621 gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg
     1681 gcaaataacc ctcgagccac ccatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     1741 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1801 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1861 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1921 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     1981 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2041 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2101 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2161 tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2221 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2281 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2341 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2401 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2461 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     2521 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     2581 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     2641 gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa
     2701 aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg
     2761 tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt
     2821 tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc
     2881 cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg ttaacgctag
     2941 catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag
//
